The present invention relates to a method for analysis of methylation of ribonucleic acid (RNA) comprising the steps: (i) contacting RNA with one or more antibodies which binds to methylated site(s) of RNA; wherein the methylated site(s) comprise at least one ribonucleotide base modified by one or more methyl groups; (ii) photo-crosslinking the one or more antibodies to crosslink individual antibodies to the RNA molecule(s) to form RNA-antibody conjugates; (iii) immunoprecipitating to separate the RNA-antibody conjugates; (iv) treating the RNA-antibody conjugates with at least one exonuclease; (v) removing the crosslinked antibodies from the RNA-antibody conjugates to release RNA; and (vi) analysing the released RNA.

The present invention relates to a method for analysis of methylation of ribonucleic acid (RNA) comprising the steps: (i) contacting RNA with one or more antibodies which binds to methylated site(s) of RNA; wherein the methylated site(s) comprise at least one ribonucleotide base modified by one or more methyl groups; (ii) photo-crosslinking the one or more antibodies to crosslink individual antibodies to the RNA molecule(s) to form RNA-antibody conjugates; (iii) immunoprecipitating to separate the RNA-antibody conjugates; (iv) treating the RNA-antibody conjugates with at least one exonuclease; (v) removing the crosslinked antibodies from the RNA-antibody conjugates to release RNA; and (vi) analysing the released RNA.

Single-cell analysis of a population of cells reveals cellular genotypes (e.g., single nucleotide variants and copy number variations) and phenotypes (e.g., protein expression) of individual cells. In one scenario, individual cells can be classified according to their respective genotypes and phenotypes. In one scenario, genotypes and phenotypes of all cells in the population are informative for identifying subpopulations of cells, thereby revealing intra-population heterogeneity. The identification of subpopulations of cells is informative for improving the understanding of cellular biology, especially in the context of diseases such as cancer, and is further informative for the better design of diagnostics and therapies.

Single-cell analysis of a population of cells reveals cellular genotypes (e.g., single nucleotide variants and copy number variations) and phenotypes (e.g., protein expression) of individual cells. In one scenario, individual cells can be classified according to their respective genotypes and phenotypes. In one scenario, genotypes and phenotypes of all cells in the population are informative for identifying subpopulations of cells, thereby revealing intra-population heterogeneity. The identification of subpopulations of cells is informative for improving the understanding of cellular biology, especially in the context of diseases such as cancer, and is further informative for the better design of diagnostics and therapies.

Presented herein are methods and compositions for enhancing specific enrichment of target sequences in a nucleic acid library. Off-target hybridization probes may be used to reduce binding and/or capture of off-target regions of a nucleic acid library in a targeted sequencing workflow. The off-target hybridization probes may be specific for locations known to generate off-target sequencing reads for a particular set of hybridization probes.

Presented herein are methods and compositions for enhancing specific enrichment of target sequences in a nucleic acid library. Off-target hybridization probes may be used to reduce binding and/or capture of off-target regions of a nucleic acid library in a targeted sequencing workflow. The off-target hybridization probes may be specific for locations known to generate off-target sequencing reads for a particular set of hybridization probes.

Provided are compositions, methods, kits, and articles of manufacture for use in RNA sequencing. The approach is referred to as easy RNA-adenylation sequencing (“Ezra-seq”). The approach provides an alternative to 3′ end linker ligation and circularization by way of an enzymatic system capable of 3′ end poly(A) tailing and 5′-end adenylation for the same RNA, using two separate enzymes, or a single fusion protein. The two enzymes or the fusion protein containing them as distinct segments are a cyclase and a polymerase. The method allows for single container processing of RNA into cDNA,

Provided are compositions, methods, kits, and articles of manufacture for use in RNA sequencing. The approach is referred to as easy RNA-adenylation sequencing (“Ezra-seq”). The approach provides an alternative to 3′ end linker ligation and circularization by way of an enzymatic system capable of 3′ end poly(A) tailing and 5′-end adenylation for the same RNA, using two separate enzymes, or a single fusion protein. The two enzymes or the fusion protein containing them as distinct segments are a cyclase and a polymerase. The method allows for single container processing of RNA into cDNA,

The present invention provides a method for detecting a plurality of analytes in a sample, comprising performing a multiplex proximity-based detection assay. The assay utilises pairs of proximity probes with shared hybridisation sites (i.e. hybridisation sites which are shared between different proximity probe pairs). Also provided is a product comprising a plurality of proximity probe pairs with shared hybridisation sites, which may be used in the method disclosed herein.

The present invention provides a method for detecting a plurality of analytes in a sample, comprising performing a multiplex proximity-based detection assay. The assay utilises pairs of proximity probes with shared hybridisation sites (i.e. hybridisation sites which are shared between different proximity probe pairs). Also provided is a product comprising a plurality of proximity probe pairs with shared hybridisation sites, which may be used in the method disclosed herein.